PMID- 37877660
OWN - NLM
STAT- MEDLINE
DCOM- 20231026
LR  - 20231027
IS  - 2045-8827 (Electronic)
IS  - 2045-8827 (Linking)
VI  - 12
IP  - 5
DP  - 2023 Oct
TI  - Isolation, characterization, and fibroblast uptake of bacterial extracellular 
      vesicles from Porphyromonas gingivalis strains.
PG  - e1388
LID - 10.1002/mbo3.1388 [doi]
LID - e1388
AB  - Periodontitis is an inflammatory condition caused by bacteria and represents a 
      serious health problem worldwide as the inflammation damages the supporting 
      tissues of the teeth and may predispose to systemic diseases. Porphyromonas 
      gingivalis is considered a keystone periodontal pathogen that releases bacterial 
      extracellular vesicles (bEVs) containing virulence factors, such as gingipains, 
      that may contribute to the pathogenesis of periodontitis. This study aimed to 
      isolate and characterize bEVs from three strains of P. gingivalis, investigate 
      putative bEV uptake into human oral fibroblasts, and determine the gingipain 
      activity of the bEVs. bEVs from three bacterial strains, ATCC 33277, A7A1-28, and 
      W83, were isolated through ultrafiltration and size-exclusion chromatography. 
      Vesicle size distribution was measured by nano-tracking analysis (NTA). 
      Transmission electron microscopy was used for bEV visualization. Flow cytometry 
      was used to detect bEVs and gingipain activity was measured with an enzyme assay 
      using a substrate specific for arg-gingipain. The uptake of bEVs into oral 
      fibroblasts was visualized using confocal microscopy. NTA showed bEV 
      concentrations from 10(8) to 10(11) particles/mL and bEV diameters from 42 to 
      356 nm. TEM pictures demonstrated vesicle-like structures. bEV-gingipains were 
      detected both by flow cytometry and enzyme assay. Fibroblasts incubated with bEVs 
      labeled with fluorescent dye displayed intracellular localization consistent with 
      bEV internalization. In conclusion, bEVs from P. gingivalis were successfully 
      isolated and characterized, and their uptake into human oral fibroblasts was 
      documented. The bEVs displayed active gingipains demonstrating their origin from 
      P. gingivalis and the potential role of bEVs in periodontitis.
CI  - (c) 2023 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.
FAU - Haugsten, Helene R
AU  - Haugsten HR
AUID- ORCID: 0009-0004-9349-3445
AD  - Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, 
      Norway.
FAU - Kristoffersen, Anne K
AU  - Kristoffersen AK
AD  - Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, 
      Norway.
FAU - Haug, Trude M
AU  - Haug TM
AUID- ORCID: 0000-0003-3776-5077
AD  - Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, 
      Norway.
FAU - Soland, Tine M
AU  - Soland TM
AD  - Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, 
      Norway.
AD  - Department of Pathology, Oslo University Hospital, Oslo, Norway.
FAU - Ovstebo, Reidun
AU  - Ovstebo R
AD  - The Blood Cell Research Group, Department of Medical Biochemistry, Oslo 
      University Hospital, Ulleval, Norway.
FAU - Aass, Hans C D
AU  - Aass HCD
AD  - The Blood Cell Research Group, Department of Medical Biochemistry, Oslo 
      University Hospital, Ulleval, Norway.
FAU - Enersen, Morten
AU  - Enersen M
AD  - Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, 
      Norway.
FAU - Galtung, Hilde K
AU  - Galtung HK
AD  - Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, 
      Norway.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Microbiologyopen
JT  - MicrobiologyOpen
JID - 101588314
RN  - 0 (Gingipain Cysteine Endopeptidases)
RN  - EC 3.4.22.- (Cysteine Endopeptidases)
RN  - 0 (Adhesins, Bacterial)
SB  - IM
MH  - Humans
MH  - Gingipain Cysteine Endopeptidases
MH  - Cysteine Endopeptidases
MH  - Porphyromonas gingivalis
MH  - Adhesins, Bacterial
MH  - *Periodontitis/microbiology
MH  - Fibroblasts/microbiology
MH  - *Extracellular Vesicles
PMC - PMC10579780
OTO - NOTNLM
OT  - Porphyromonas gingivalis
OT  - bacterial vesicle
OT  - virulence factors
COIS- None declared.
EDAT- 2023/10/25 12:42
MHDA- 2023/10/26 06:42
PMCR- 2023/10/16
CRDT- 2023/10/25 08:38
PHST- 2023/10/04 00:00 [revised]
PHST- 2023/06/15 00:00 [received]
PHST- 2023/10/06 00:00 [accepted]
PHST- 2023/10/26 06:42 [medline]
PHST- 2023/10/25 12:42 [pubmed]
PHST- 2023/10/25 08:38 [entrez]
PHST- 2023/10/16 00:00 [pmc-release]
AID - MBO31388 [pii]
AID - 10.1002/mbo3.1388 [doi]
PST - ppublish
SO  - Microbiologyopen. 2023 Oct;12(5):e1388. doi: 10.1002/mbo3.1388.